The families Violaceae (alternatively known as Alsodeiace or Leoniaceae or Retrosepalaceae) comprise twenty genera and about 800 species. Viola serpens belongs to family Violaceae and commonly known as “Banafsa”. It is a small glabrous, perennial herb, which is found throughout India in moist woods and hilly districts. The essential oil of aerial parts of V. serpens, were extracted by steam distillation. The quantitative and qualitative analysis of volatile essential oil constituents of the plant was done by Gas Chromatography (GC) and GC-Mass Spectrometry. A total of 50 components of the essential oil of V. serpens were identified, accounting for 81.38% of the total oil. The main compounds found were Bis (2- ethylhexyl) maleate (15.62%), 2, 4, 4, 6-Tetramethyl-2-heptene (11.52%), Hexen-3-ol (6.56%), and Cis Verbeno (l 4.77%). The chemical constituents in the essential oil from V. serpens were identified in the following classes or groups of chemical compounds, such as monoterpens, sesquiterpenes volatile organic compounds and their oxygenated hydrocarbons. Therefore, the essential constituents could be used as antioxidant, antifungal or antimicrobial agent in new drugs preparation for therapy of infectious diseases.